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H Mullink

Publications and source records attributed to H Mullink.

At least 19 recordsLinked to original sources
PubMed1992

Epstein-Barr virus DNA in Reed-Sternberg cells of Hodgkin's disease is frequently associated with CR2 (EBV receptor) expression.

We studied 44 cases of Hodgkin's disease for the presence of Epstein-Barr virus (EBV) DNA, its localization and the expression of the EBV receptor on the tumour cells. EBV DNA was found in 52% (16/31) of the Hodgkin's lymphomas using the polymerase chain reaction. With a very sensitive non-radioactive DNA in situ hybridization technique in combination with immunohistochemistry for CD 30 or CD 15 antigens, EBV DNA was localized to Reed-Sternberg cells and its mononuclear variants. The relationship between the presence of EBV DNA and the expression of the EBV-receptor CR2 (CD 21) on Reed-Sternberg cells was studied using the same techniques and two different monoclonal anti-CD 21 antibodies. CR2 could be detected on a substantial number of the Reed-Sternberg cells in EBV DNA positive Hodgkin's lymphomas (9/12; 75%), whereas in EBV negative cases positivity with anti-CD 21 was rare (1/13; 8%). The results indicate that CR2 expression on Reed-Sternberg cells and the presence of EBV DNA sequences are frequently associated in Hodgkin's lymphomas.

Antigens, Differentiation, B-Lymphocyte↗
PubMed1992

Application and comparison of silver intensification methods for the diaminobenzidine and diaminobenzidine-nickel endproduct of the peroxidation reaction in immunohistochemistry and in situ hybridization.

Silver-intensification methods described in the literature for the diaminobenzidine (DAB) and diaminobenzidine-nickel (DAB/Ni) endproduct of the peroxidase reaction were compared in model systems after immunoperoxidase and in situ hybridization. First, these methods were compared in immunohistochemical model systems, using the demonstration of glial fibrillar acidic protein (GFAP) and prostate-specific antigen (PSA) in paraffin sections of human brain and prostate tissue, respectively. When DAB without Ni was used as substrate, tissue argyrophilia caused considerable background staining. Only when this tissue reactivity was quenched with, e.g., CuSO4 with H2O2 or thioglycolic acid, were the results acceptable. A considerable improvement in the signal-to-noise ratio could be obtained when nickel was included in the substrate mixture. The methods that proved to be best for demonstration of GFAP and PSA made use of acid developer solutions. Subsequently, these methods were compared with other sensitive immunostaining methods for demonstration of the gamma-delta T-cell receptor in frozen lymphoid tissue. In this model a considerable increase in the number of positive cells could be obtained using silver intensification. The different methods using DAB/Ni were also compared for use in DNA in situ hybridization (DISH). In this case two model systems were used: human papilloma virus type 11 (HPV-11) DNA in condyloma tissue (abundant target model) and Epstein-Barr virus (EBV) DNA in a mononucleosis lymph node (low target model). For demonstration of HPV-11, all methods gave more or less satisfactory results, which were best with the acid developer solutions. Moreover, for demonstration of EBV DNA, a signal could be obtained only with these developer solutions. Such a method also proved suitable in double immuno-hybrido stainings for the demonstration of EBV DNA in specific antigen-positive Reed-Sternberg cells in paraffin sections of Hodgkin lymph nodes.

3,3'-Diaminobenzidine↗
PubMed1991

Oral hairy leukoplakia in HIV infection: a diagnostic pitfall.

Twenty-nine human immunodeficiency virus (HIV)-infected patients with white, nonremovable lesions on the lateral border of the tongue, clinically suggestive of oral hairy leukoplakia (HL), were studied. In particular, the value of local antifungal therapy in establishing the diagnosis of HL was investigated. In 15 patients (52%) the lesions could be ultimately attributed to a candidal infection of the tongue. In 10 of the remaining 14 patients, a biopsy was obtained from lesions persisting after local antifungal treatment. In all biopsy specimens, the diagnosis of HL was confirmed by histopathologic examination and the demonstration of Epstein-Barr virus DNA by polymerase chain reaction, Southern blot hybridization, and DNA in situ hybridization. The present data confirm that the diagnosis of HL in HIV-infected patients cannot be reliably made on clinical criteria alone, but requires histopathologic confirmation including the demonstration of Epstein-Barr virus DNA, preferably by DNA in situ hybridization. However, with regard to the differential diagnosis of white, nonremovable lesions on the lateral border of the tongue in HIV-infected patients, the present study suggests that persistence of lesions after local antifungal therapy is highly suggestive of HL.

Adolescent↗
PubMed1991

Demonstration of changes in cytokeratin expression in condylomata accuminata in relation to the presence of human papilloma virus as shown by a combination of immunohistochemistry and in situ hybridization.

Human papillomavirus (HPV) can be detected in, and is probably involved in the etiology of, the majority of anogenital neoplasias. Infection with the virus induces a number of events in the infected epithelial cells that may lead to the development of benign or malignant tumors. One change that can be detected in the infected cells is in squamous differentiation, which is reflected by the pattern of cytokeratin polypeptide expression. By studying this pattern in relation to the presence of the virus, an indication may be obtained of the influence of the virus on the cellular differentiation in individual cells. By using a combination of DNA in situ hybridization and immunohistochemistry, for HPV and cytokeratin polypeptides, respectively, we studied the presence of HPV6 or HPV11 in condylomata accuminata derived from anogenital skin in relation to the cytokeratin polypeptides K1, 4, 8, 10, 14, and 18. We found that in many samples the presence of the skin-type cytokeratins K1 and K10 was decreased, whereas K13, and to a lesser degree K4, appeared. The cellular localization of these aberrations in cytokeratin expression could be related to the presence of HPV6 or 11 DNA in the tissue.

Cell Differentiation↗
PubMed1991

Macrophage-T suppressor cell interference in the lungs of steroid-treated sarcoidosis patients.

In the bronchoalveolar lavage of sarcoidosis patients the mononuclear cell infiltrate was enumerated on T helper and suppressor lymphocytes as well as macrophages by means of a triple-staining assay on cytospin slides. As was seen on the slides, lymphocytes were often adhered very closely to macrophages. This phenomenon, many times described but not understood, was studied in a group of 13 sarcoidosis patients, of whom 7 received prednisolone treatment. It could be shown that treatment with the corticosteroid was followed by an increase in the percentage of suppressor lymphocytes adhered to macrophages. Second, the number of such alveolar T suppressor lymphocyte-macrophage aggregates was dramatically increased in the prednisolone-treated patients.

Adult↗
PubMed1990

Detection of human papillomavirus and Epstein-Barr virus DNA sequences in oral mucosa of HIV-infected patients by the polymerase chain reaction.

The presence of human papillomavirus (HPV) and Epstein-Barr virus (EBV) was analyzed in 21 oral biopsy specimens of HIV-infected patients using the polymerase chain reaction (PCR) method. Biopsies were categorized as hairy leukoplakia (HL) (n = 12), candidiasis (n = 3), oral warts (n = 2), and clinically normal epithelium (n = 4). For HPV detection a modified general primer-mediated PCR method (GP-PCR), which detects a broad spectrum of HPV genotypes at sub-picogram levels, was used. Human papillomavirus DNA was only found in two oral warts and was identified as HPV type 32. Epstein-Barr virus DNA was detected in 16 biopsy specimens, including the 12 HLs, 2 cases of candidiasis, and 2 samples of normal epithelium. Epstein-Barr virus positivity in HL could be confirmed by Southern blot analysis and DNA in situ hybridization using biotinylated DNA probes (bio-DISH). Epstein-Barr virus bio-DISH was also positive in one sample of normal epithelium from a patient with HL. The results indicate that HL is strongly associated with EBV and not with any of the common HPV types that react with general HPV primers in the PCR. However the detection of EBV in normal oral epithelium by PCR and bio-DISH suggests that the presence of this virus is not exclusively related to HL.

Base Sequence↗
PubMed1989

Two colour DNA in situ hybridization for the detection of two viral genomes using non-radioactive probes.

Methods for the simultaneous detection of two virus types in cytological preparations or tissue sections by non-radioactive in situ hybridization were investigated. As a model system, CaSki cells, which have human papilloma virus type 16 (HPV16) DNA integrated in their cellular genome, were in vitro infected with Herpes simplex virus 2 (HSV2). DNA probes for both viruses were labeled with biotin, acetylaminofluorene (AAF), and transaminated-sulfonated cytosine (TS-modified). Best results were obtained when a mixture of biotinated and haptenized DNA probes (AAF- or TS-modified) was used for hybridization. The biotinated hybrid was demonstrated with a streptavidin-biotinated alkaline phosphatase staining reaction, whereas the haptenized hybrid was visualized by an indirect peroxidase method. Visualisation of both viral DNAs in the same cell was possible by a combination of biotinated HPV16 DNA and haptenized HSV2 DNA.

DNA Probes, HPV↗
PubMed1989

Combined immuno- and non-radioactive hybridocytochemistry on cells and tissue sections: influence of fixation, enzyme pre-treatment, and choice of chromogen on detection of antigen and DNA sequences.

Conditions for combination of DNA in situ hybridization, using biotinylated DNA probes, with immunohistochemistry were investigated on cryostat sections, cytological preparations, and paraffin sections. We found that cryostat sections and cytological preparations are suitable for in situ hybridization of target DNA after fixation in acetone, methanol, ethanol, or Carnoy without further proteinase pretreatment. Acetone is also very suitable for immunostaining of cell surface or cytoskeleton antigens. We therefore performed combined immunoenzyme and in situ hybridization staining using this fixative. The best results were obtained when immunoperoxidase staining with diaminobenzidine/H2O2 was followed directly by in situ hybridization. In addition to immunoperoxidase, alkaline phosphatase-antialkaline phosphatase (APAAP) staining with naphthol ASBI phosphate and New Fuchsin as a substrate could be used. In most instances, detection of the biotinylated hybrid with a streptavidin-biotinylated polyalkaline phosphatase method using nitroblue tetrazolium and 5-bromo-4-chloro-3-indolylphosphate as the substrate was preferable. The double stainings were studied on the following test models: (a) frozen tonsil sections: cell surface antigens (pan T) and ribosomal DNA; (b) frozen genital condyloma sections; cytokeratins and human papillomavirus type 6 + 11 (HPV-6/11) DNA; (c) CaSKi cells: cytokeratins and HPV-16 DNA; (d) infected fetal lung fibroblasts: vimentin and cytomegalovirus (CMV) DNA. An adapted procedure was followed on routinely formaldehye-fixed and paraffin-embedded condyloma tissue. Immunoperoxidase staining for papilloma virus capsid antigen could be combined with DNA in situ hybridization with HPV-6/11 DNA. In this model, however, the accessibility of the target DNA had to be improved by enzyme treatment after the immunostaining and before starting the in situ hybridization.

Antigens, Surface↗
PubMed1989

Immunohistochemistry in bone marrow diagnosis. Value of a panel of monoclonal antibodies on routinely processed bone marrow biopsies.

To see if immunohistochemistry can be used on routinely processed bone marrow biopsies for diagnostic purposes, 73 biopsy specimens, fixed in sublimate-formaldehyde, decalcified in an acetic acid-formaldehyde mixture, and embedded in paraffin, were studied with a panel of antibodies. The specimens included "normal," lymphomatous, and myeloproliferative disorders as well as some biopsies with metastatic carcinoma. The results show that the different cell lines and their localization in the bone marrow can be easily identified and quantitative and qualitative changes can be assessed. Megakaryopoiesis is identified with Factor VIII-related antigen and Ulex europaeus agglutinin (UEA); myelopoiesis stains with MT-1, elastase, Leu M-1, LN-2, LN-3, HECA 452, and 115D8; and in myeloproliferative conditions, myeloblasts and promyelocytes stained with leukocyte common antigen (LCA). Erythroid cells stained with UEA, glycophorin A, and LN-1. Lymphocytes were marked with LCA, MB-2, and LN-2. Plasma cells were stained best with immunoglobulin light chain antisera; only occasional reactivity with LCA and 115D8 was observed. Carcinomas all reacted with MB-2; occasional reactivity with 115D8, HECA 452, LN-1, LN-2, MT-1, and Ber H-2 was seen. A small panel of selected antibodies, such as UEA, Leu M-1, and LCA, and the immunoglobulin light chain antisera can be very helpful in bone marrow diagnosis and would cover most indications.

Antibodies, Monoclonal↗
PubMed1988

Diagnostic application of panels of antibodies in mucosal melanomas of the head and neck.

This article describes the use of panels of antibodies in the histopathologic diagnosis of ten malignant melanomas arising in the mucosa of the head and neck. The immunohistochemistry analysis was performed in a step-by-step manner. In the first step a panel of antibodies discriminating between carcinoma, malignant lymphoma, and melanoma was used. Staining with antivimentin and the melanoma-associated antigen, moAb NKI/C-3, was found to occur in all cases. The NKI/C-3 stained nine of 10 specimens from the first biopsy. Anti-S-100 protein, anti-HMW-MAA, and NKI-beteb was used as a panel in the second step. These stained ten of ten, five of six, and seven of seven cases, respectively. The diagnosis for melanin was confirmed by extensive sampling and a diligent search for melanin and (pre)melanosomes in different biopsy and surgical specimens. These results indicate that a step-wise application of immunohistochemistry is a very useful tool in the diagnosis of mucosal melanoma of the head and neck.

Aged↗
PubMed1988

Immunohistochemical localization of pepsinogen A and C containing cells in Barrett's oesophagus.

No data are available on the localization of Pepsinogen A (PGA = PG I) and Pepsinogen C (PGC = PG II) positive cells in Barrett's epithelium. Endoscopic biopsy specimens were taken from the columnar epithelium from 23 patients (n = 93), and in addition from the cardia from eight healthy control subjects (n = 38). The tissue was stained by the immunoperoxidase technique with specific anti-pepsinogen antisera, and double immunostained for PGA and PGC. In the Barrett's epithelium PGA was found in 28 out of 93 biopsy specimens (30.1%) and PGC in 55 out of 93 (59.1%). Chief cells always stained both for PGA- and PGC +. PGA + and PGC + cells were found each in 100% of the biopsy specimens with fundic type epithelium, in 21.7% and 70.7% of biopsy specimens with junctional type, in 0% and 26.1% of biopsy specimens with specialized epithelium and in 12.5% and 43.5% of biopsy specimens with mixed junctional/specialized features respectively. Dysplastic epithelium stained always negatively with both anti-pepsinogen antisera. In most control cardia biopsy specimens PGA as well as PGC were demonstrable; occasionally clear mucous glands were PGA - and PGC+. It is concluded that pepsinogen-containing cells can be accurately identified in the Barrett's epithelium; their presence seems related to the histological cell type. Identification of pepsinogen positive cells may contribute to a more accurate morphological classification of the Barrett's epithelium.

Adolescent↗
PubMed1988

Expression of intermediate filament proteins in adrenal cortex and related tumours.

The intermediate filament profile of adrenal cortex and its related tumours has been evaluated. Most adrenocortical cells contained cytokeratin 8 and 18 as demonstrated by monoclonal antibodies CAM 5.2, M20, M9 and RGE53. Cytokeratin immunoreactivity was not confined to a functional zone of the adrenal cortex. Only a small number of the adrenocortical cells showed vimentin immunoreactivity. From normal adrenal cortex through adenomas, to carcinomas, there is a progressive decrease or even loss of cytokeratin immunoreactivity and an increase in vimentin immunoreactivity. Aberrant cytokeratin expression was not found in adrenocortical adenomas and carcinomas with the antibodies used. Awareness of the possible absence of cytokeratin immunoreactivity in adrenocortical carcinomas is important whenever antibodies to cytokeratins and vimentin are used for diagnostic purposes in poorly differentiated neoplasms.

Adenoma↗
PubMed1988

The expression of intermediate filaments and mam-6 antigen in relation to the degree of morphologic differentiation of carcinoma of the head and neck: diagnostic implications.

In this study, the immunoreactivity of several cytokeratin antibodies; 115 D8, a monoclonal antibody against MAM-6, an epithelial membrane antigen; and two vimentin antibodies, is examined in relation to the degree of morphologic differentiation in carcinomas of the head and neck. The results indicate that a relationship exists between the degree of morphologic differentiation and the expression of cytokeratin, MAM-6, and vimentin, as detected by polyclonal antikeratin, 115 D8 and anti-vimentin. Expression of cytokeratin and MAM-6 is reversely related to vimentin. Polyclonal anti-keratin; CAM 5.2, a monoclonal antibody against cytokeratin 8, 18 and 19; and 115 D8, used in combination, were still able to identify the epithelial nature of undifferentiated/spindle cells. Since these immunohistochemical markers precede light microscopic detectable signs of epithelial differentiation, they can be used for the identification of the epithelial nature of undifferentiated/spindle tumors of the head and neck.

Antigens, Differentiation↗
PubMed1988

Sensitivity of in situ detection with biotinylated probes of human papilloma virus type 16 DNA in frozen tissue sections of squamous cell carcinomas of the cervix.

The sensitivity of human papilloma virus type 16 (HPV-16) DNA detection by DNA in situ hybridization using biotinylated probes (bio-DISH) was estimated by performing this technique on snap-frozen tissue sections of 10 cervical squamous cell carcinomas containing increasing amounts of HPV-16 as determined by Southern blot hybridization. A protocol using serial sections for bio-DISH and DNA extraction was used. The number of positively stained cells and the detection limit were strongly dependent on the treatment of the sections with proteinase K prior to hybridization. At low proteinase K concentration (0.1 micrograms/ml), the detection limit appeared to be 30-40 HPV-16 DNA copies per carcinoma cell, whereas morphology was preserved. A high proteinase K concentration (1-5 micrograms/ml) often resulted in an increase in the number of positively stained cells but also in a poor morphology. The detection limit was improved to at least 20 HPV-16 DNA copies per carcinoma cell.

Biotin↗
PubMed1987

A rapid and simple hapten conjugation method for monoclonal antibodies to be used in immunoenzyme single and double staining procedures.

A rapid and simple method was developed for the haptenization of monoclonal antibodies (MAbs), to be used in immunoenzyme single and double staining techniques. Using this method minute amounts of MAbs can be haptenized without purification of the antibody or removal of the excess hapten. The haptenized MAb can be ready for use with 2.5 h. The method consists of a direct incubation of the antibody with the haptenizing agent and subsequent addition of an amino acid solution to stop the reaction. Commonly available reagents were tested, of which dinitrofluorobenzene, trinitrobenzene sulfonic acid and an oxazolone derivative gave the best results. The procedure was evaluated by using MAbs directed against lymphoid cell surface membrane antigens in an indirect immunoenzyme staining on frozen sections using peroxidase or alkaline phosphatase-conjugated anti-hapten antibodies as second step antibody. It was found that those MAbs, which show good staining results in conventional indirect immunoenzyme procedures, can also be used successfully after haptenization in single as well as double staining procedures. Combination of haptenized and biotynilated MAbs gave good results when weakly reactive MAbs had to be included in immunoenzyme double staining.

Antibodies, Monoclonal↗
PubMed1987

Expression of cytokeratins and vimentin in epithelial cells of normal and pathologic thyroid tissue.

The presence of intermediate filament proteins of the cytokeratin and vimentin type was evaluated in normal and pathologically changed thyroid tissue specimens. Using the indirect immunoperoxidase technique with 4 different cytokeratin monoclonal antibodies: RCK114 (broad spectred), K2080 (broad spectred), RGE53 (directed against component 18, present in simple epithelium) and RKSE60 (directed against component 10, associated with keratinization). Co-expression of cytokeratin and vimentin was evaluated with a double immunoenzyme staining technique. The results indicate that normal and transformed cells express cytokeratins of the non-epidermal type. Cytokeratins of the epidermal type are sometimes present in carcinomas. They do not differentiate in tumour type (i.e. papillary, follicular, anaplastic or medullary carcinoma). The co-expression of cytokeratins and vimentin is not restricted to carcinomas: in a small percentage of cases it is also present in normal epithelial cells of the thyroid gland. Moreover, the distribution pattern of cytokeratins and vimentin within the cell is changed in malignant transformed epithelial cells of the gland and seems to be inversely related to the degree of differentiation of these cells. The implications of our findings for the possible use of cytokeratins and vimentin in diagnostic pathology are discussed.

Antibodies, Monoclonal↗